Nebivolol Polymeric Nanoparticles-Loaded In Situ Gel for Effective Treatment of Glaucoma: Optimization, Physicochemical Characterization, and Pharmacokinetic and Pharmacodynamic Evaluation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Analytical and Bioanalytical Methods
2.3. Selection of Excipients and Process Parameters for the Preparation of NEB-PNPs
2.4. Formulation of NEB-PNPs
2.4.1. Optimization by Applying Design of Experiments (DoE) for the Preparation of NEB-PNPs
2.4.2. Optimization and Validation of the Regression Equations
2.5. Evaluation of Various Physical Characteristics of NEB-PNPs
2.5.1. Analysis of Particle Size and Zeta Potential of NEB-PNPs
2.5.2. Analysis of Drug Loading and Entrapment Efficiency of NEB-PNPs
2.6. Scanning Electron Microscopy (SEM) Imaging of the Optimized NEB-PNPs
2.7. Differential Scanning Calorimetry (DSC) of the Optimized NEB-PNPs
2.8. X-ray Diffractometric Analysis of the Optimized NEB-PNPs
2.9. Preparation of NEB-PNPs-Loaded Dual-Sensitive In Situ Gel
2.10. Rheological Evaluation of NEB-PNPs-ISG Formulation
2.11. In Vitro Drug-Release Studies of NEB-PNPs-Susp and NEB-PNPs-ISG Formulations
2.12. In Vivo Evaluation of Optimized NEB-PNPs and NEB-PNPs-ISG Formulations
2.12.1. Ocular Pharmacokinetic Studies
2.12.2. Ocular Pharmacodynamic Studies of NEB-PNP Formulations
3. Results and Discussion
3.1. Preliminary Trials for the Preparation of NEB-PNPs
3.2. Optimization of NEB-PNPs
3.2.1. Impact of Critical Factors on Particle Size of NEB-PNPs
3.2.2. Impact of Critical Factors on Drug Loading of NEB-PNPs
3.2.3. Validation of the Regression Equations for Particle Size and Loading Efficiency
3.3. Physical Characterization of the Optimized NEB-PNPs Using Zeta-Sizer, SEM, DSC, and pXRD
3.4. Rheological Evaluation of NEB-PNPs-ISG Formulation
3.5. Drug-Release Studies of NEB-PNPs Formulations
3.6. Stability Studies of PNPs Formulations
3.7. In Vivo Studies of the NEB-PNPs Formulations
3.7.1. Ocular Pharmacokinetic Studies of NEB-PNPs Formulations
3.7.2. Ocular Pharmacodynamic Studies
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Design used for optimization | Box–Behnken design (BBD): A response surface methodology suitable to optimize and determine the mathematical equations relating three or four independent factors and one or more critical responses. |
Criteria for selection the design | Though BBD and central composite design (CCD) are the two most popular optimization designs available, BBD was selected for the following reasons:
|
No. of experimental runs used in the design | In the current work, which involves three critical factors, a total 17 runs were performed for optimization (12 factorial runs + 5 center-point runs) |
Critical factors | X1—amount of PCL; X2—concentration of stabilizer; X3—homogenization speed |
Critical responses | Y1—particle size; Y2—drug loading (%) |
Method to determine optimal solution | Simultaneous optimization of both the critical responses based on the highest overall desirability value. |
Source | Particle Size (Y1) | Drug Loading (Y2) | ||||||
---|---|---|---|---|---|---|---|---|
SS | DF | Fcal | Pcal | SS | DF | Fcal | Pcal | |
Model | 38,605.75 | 3 | 36.68 | <0.0001 | 0.018 | 1 | 37,300 | <0.0001 |
X1 | 27,097.92 | 1 | 77.23 | <0.0001 | 0.018 | 1 | 37,300 | <0.0001 |
X3 | 8508.6 | 1 | 24.25 | 0.0003 | ||||
X32 | 2999.23 | 1 | 8.55 | 0.0119 | ||||
Residual | 4561.33 | 13 | 0.00001 | 15 | ||||
Lack-of-Fit | 3913.55 | 9 | 2.69 | 0.1773 | 0.0001 | 11 | 1.16 | 0.482 |
Pure Error | 647.78 | 4 | 0.000002 | 4 | ||||
Total | 43,167.08 | 16 | 0.0177 | 16 |
Composition of Optimized NEB-PNPs | * Physicochemical Characteristics of Optimized NEB-PNPs | ||||
---|---|---|---|---|---|
PS (nm) | PDI | ZP (mV) | EE (%) | DL (%) | |
Organic Phase: 10 mg of NEB + 25 mg of PCL dissolved in 1 mL of NMP Aqueous Phase: PVA (0.75% w/v in water) | 270.9 ± 6.3 | 0.24 ± 0.03 | −8.2 ± 1.2 | 96.7 ± 0.3 | 28.8 ± 2.4 |
Biological Matrix | PK Parameters | Units | Treatments | ||
---|---|---|---|---|---|
NEB-Susp # | NEB-PNPs-Susp | NEB-PNPs-ISG | |||
Aqueous humor | Cmax a | ng/mL | 28.2 ± 3.1 | 36.8 ± 3.2 | 30.2 ± 2.1 |
Tmax b | h | 2.0 | 4.0 | 4.0 | |
AUC0–t c | ng × h/mL | 189 | 204.4 | 329.2 | |
MRT0–∞ c | h | 6.1 | 6.4 | 9.7 | |
Plasma d | Cmax | ng/mL | 1.86 ± 0.1 | 1.15 ± 0.08 | 0.58 ± 0.03 |
Tmax | h | 1.0 | 2.0 | 4.0 | |
AUC0–t | ng × h/mL | 20.2 ± 2.7 | 12.1 ± 0.9 | 8.38 ± 0.56 | |
MRT0–∞ | h | 25.8 ± 1.5 | 10.4 ± 1.1 | 4.6 ± 0.4 |
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Rawat, P.S.; Ravi, P.R.; Khan, M.S.; Mahajan, R.R.; Szeleszczuk, Ł. Nebivolol Polymeric Nanoparticles-Loaded In Situ Gel for Effective Treatment of Glaucoma: Optimization, Physicochemical Characterization, and Pharmacokinetic and Pharmacodynamic Evaluation. Nanomaterials 2024, 14, 1347. https://doi.org/10.3390/nano14161347
Rawat PS, Ravi PR, Khan MS, Mahajan RR, Szeleszczuk Ł. Nebivolol Polymeric Nanoparticles-Loaded In Situ Gel for Effective Treatment of Glaucoma: Optimization, Physicochemical Characterization, and Pharmacokinetic and Pharmacodynamic Evaluation. Nanomaterials. 2024; 14(16):1347. https://doi.org/10.3390/nano14161347
Chicago/Turabian StyleRawat, Pradeep Singh, Punna Rao Ravi, Mohammed Shareef Khan, Radhika Rajiv Mahajan, and Łukasz Szeleszczuk. 2024. "Nebivolol Polymeric Nanoparticles-Loaded In Situ Gel for Effective Treatment of Glaucoma: Optimization, Physicochemical Characterization, and Pharmacokinetic and Pharmacodynamic Evaluation" Nanomaterials 14, no. 16: 1347. https://doi.org/10.3390/nano14161347